• Title/Summary/Keyword: Liquid membrane

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Primary Culture of Human Nasal Epithelial Cell Monolayer for In Vitro Drug Transport Studies (약물의 in vitro 투과 실험을 위한 사람의 비강점막상피세포 단층막의 일차배양)

  • Yoo, Jin-Wook;Kim, Yoo-Sun;Lee, Min-Ki;Roh, Hwan-Jung;Lee, Chi-Ho;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.32 no.1
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    • pp.21-26
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    • 2002
  • The primary culture of human nasal epithelial cell monolayer was performed on a Transwell. The effect of various factors on the tight junction formation was observed in order to develop an in vitro experimental system for nasal transport studies. Human nasal epithelial cells, collected from human normal inferior turbinates, were plated onto diverse inserts. After 4 days, media of the apical surface was removed for air-liquid interface (ALI) culture. Morphological characteristics was observed by transmission electron microscopy (TEM). A polyester membrane of $0.4\;{\mu}m$ pore size was determined as the most effective insert based on the change in the transepithelial electric resistance (TEER) value as well as the $^{14}C-mannitol$ transport study. The ALI method was effective in developing the tight junction as observed in the further increase in the TEER value and reduction in the permeability coefficient $(P_{app})$ of $^{14}C-mannitol$ transport. Results of the transport study of a model drug, budesonide, showed that the primary culture system developed in this study could be further developed and applied for in vitro nasal transport studies.

Effects of Different Concentrations of Escherichia coli and Days of Preservation on Boar Sperm Quality

  • Chung, Ki-Hwa;Kim, In-Cheul;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.213-217
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    • 2013
  • The objective of this study was to investigate the effect of bacterial contamination on elapsed time after preservation on boar semen. Known numbers of Escherichia coli (E. coli) were inoculated to freshly ejaculated semen and sperm parameters such as viability, motility, agglutination, acrosome integrity and hypo-osmotic swelling test were performed during 7 days of liquid preservation. Semen samples were prepared using antibiotic free BTS extender and 4 different levels of E. coli were treated to semen with following concentrations; 3,000, 5,000, 7,000, 10,000 CFU/ml of sperms. Semen samples were preserved at $17^{\circ}C$ for 7 days in semen storage until analyzed. Aliquots were subjected to measure the sperm viability, motility and agglutination using computer assisted sperm analysis (CASA) system, acrosome integrity was performed using chlortetracycline (CTC) staining method and hypo-osmotic swelling test was performed using hypotonic solution from day 1 (day of semen collection) to 7. Detrimental effects on sperm motility and viability were observed 3 days after preservation at the level of 5,000 CFU/ml (p<0.05). Percentage of sperm abnormality was higher (p<0.05) in over 5,000 CFU/ml groups. Sperm agglutination rate was also significantly higher (p<0.05) in groups of 5,000 and 7,000 CFU/ml. The rate of acrosome reacted sperm was higher as preservation time goes in all the samples but the pattern was clearly higher among E. coli contaminated groups (p<0.05). The sperm membrane integrity in terms of hypo-osmotic test, E. coli affects little compared to other sperm parameters. The deleterious effects observed due to the bacterial contamination in semen suggest that importance of hygiene protocol to minimize the bacterial contamination during semen collection and processing.

Simulation of Ice Ring Formation around Cryogenic Underground Storage Cavern using Hydro-Thermal Coupling Method (극저온 지하저장고 주변 ice ring 생성 모델링을 위한 열-수리 해석)

  • Jung Yong-Bok;Park Chan;Chung So-Keul;Jeong Woo-Cheol;Kim Ho-Yeong
    • Tunnel and Underground Space
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    • v.16 no.3 s.62
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    • pp.241-250
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    • 2006
  • Ice ring formation, one of the core techniques in LNG storage in a lined rock cavern, is investigated through hydro-thermal coupled analysis. An ice ring acts as a secondary barrier in case of leakage of cryogenic liquid and as a primary barrier for groundwater intrusion into an LNG cavern. Therefore, the thickness and location of the ice ring are crucial factors for the safe operation of an LNG storage cavern, especially for maintaining the integrity of a primary barrier composed of concrete, PU foam, and steel membrane. Through numerical analyses, the position and thickness of the ice ring are estimated, and the temperature and groundwater level are compared with measured values. The temperature md groundwater level by numerical analyses show good agreement with the field measurements when temperature-dependent properties and phase change are taken into account. The schemes used in this paper can be applied for estimation of ice ring formation in designing a full-scale LNG cavern.

Expression and Purification of Transmembrane Protein MerE from Mercury-Resistant Bacillus cereus

  • Amin, Aatif;Sarwar, Arslan;Saleem, Mushtaq A.;Latif, Zakia;Opella, Stanley J.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.274-282
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    • 2019
  • Mercury-resistant ($Hg^R$) bacteria were isolated from heavy metal polluted wastewater and soil collected near to tanneries of district Kasur, Pakistan. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to $40{\mu}g/ml$ against mercuric chloride ($HgCl_2$). 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of selected isolates as Bacillus sp. AZ-1 (KT270477), Bacillus cereus AZ-2 (KT270478) and Bacillus cereus AZ-3 (KT270479). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury-resistant Gram-positive bacteria. The merE gene involved in the transportation of elemental mercury ($Hg^0$) via cell membrane was cloned for the first time into pHLV vector and transformed in overexpressed C43(DE3) E. coli cells. The recombinant plasmid (pHLMerE) was expressed and the native MerE protein was obtained after thrombin cleavage by size exclusion chromatography (SEC). The purification of fusion/recombinant and native protein MerE by Ni-NTA column, dialysis and fast protein liquid chromatography (FPLC/SEC) involved unfolding/refolding techniques. A small-scale reservoir of wastewater containing $30{\mu}g/ml$ of $HgCl_2$ was designed to check the detoxification ability of selected strains. It resulted in 83% detoxification of mercury by B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 respectively (p < 0.05).

Effect of green tea extract in extender of Simmental bull semen on pregnancy rate of recipients

  • Susilowati, Suherni;Sardjito, Trilas;Mustofa, Imam;Widodo, Oky Setio;Kurnijasanti, Rochmah
    • Animal Bioscience
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    • v.34 no.2
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    • pp.198-204
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    • 2021
  • Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows. Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination. Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12). Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

Analysis of the current status and implications of nitrogen recovery from livestock manure (가축분뇨로부터 질소 회수 연구 현황 및 시사점 분석)

  • Im, Seongwon;Kim, Sangmi;Kim, Jimin;Kim, Dong-Hoon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.29 no.1
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    • pp.37-46
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    • 2021
  • Nitrogen and phosphorus in livestock manure are environmental pollutants, but also could be valuable industrial resources. In the present study, we (1) introduced various nitrogen removal technologies such as stripping, thermal method, membrane, and electrodialysis, (2) reviewed relevant studies reported in 2011-2020, in particular, full-scale experiences, and (3) assessed each technologies based on the above survey results. In addition, we provided the information on the appropriate range of the pH, temperature, gas and liquid ratio, and so on in ammonia stripping process, and expected mass balance when it is connected to biogasification process. We hope the content herein can be helpful for making policy and operating full-scale plant in Korea.

Profiling of differentially expressed proteins between fresh and frozen-thawed Duroc boar semen using ProteinChip CM10

  • Yong-Min Kim;Sung-Woo Park;Mi-Jin Lee;Da-Yeon Jeon;Su-Jin Sa;Yong-Dae Jeong;Ha-Seung Seong;Jung-Woo Choi;Shinichi, Hochi;Eun-Seok Cho;Hak-Jae Chung
    • Journal of Animal Science and Technology
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    • v.65 no.2
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    • pp.401-411
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    • 2023
  • Many studies have been conducted to improve technology for semen cryopreservation in pigs. However, computer-assisted analysis of sperm motility and morphology is insufficient to predict the molecular function of frozen-thawed semen. More accurate expression patterns of boar sperm proteins may be derived using the isobaric tags for relative and absolute quantification (iTRAQ) technique. In this study, the iTRAQ-labeling system was coupled with liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis to identify differentially expressed CM10-fractionated proteins between fresh and frozen-thawed boar semen. A total of 76 protein types were identified to be differentially expressed, among which 9 and 67 proteins showed higher and lower expression in frozen-thawed than in fresh sperm samples, respectively. The classified functions of these proteins included oxidative phosphorylation, mitochondrial inner membrane and matrix, and pyruvate metabolic processes, which are involved in adenosine triphosphate (ATP) synthesis; and sperm flagellum and motile cilium, which are involved in sperm tail structure. These results suggest a possible network of biomarkers associated with survival after the cryopreservation of Duroc boar semen.

A Study on Optimal Operation of Methanol Steam Reforming System for Hydrogen Fuel Cell Propulsion Ships (수소 연료전지 추진 선박 적용을 위한 메탄올 수증기 개질 시스템 최적 운전점 연구)

  • HEEJOO CHO;SOOBIN HYEON;SEUNGKYO JUNG;HYUNJIN JI;JUNGHO CHOI
    • Journal of Hydrogen and New Energy
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    • v.33 no.6
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    • pp.733-742
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    • 2022
  • Hydrogen fuel cell propulsion ships are emerging to respond to the recently strengthened carbon emission regulations in the international shipping sector. Methanol can be stored in a liquid state at normal pressure and temperature, and has the advantage of lower reforming temperature compared to other fuels. In this study, the optimal operating point of the methanol steam reforming system was derived by changing the Steam Carbon Ratio (SCR) from 0.10 to 3.00. Results showed that In terms of methanol conversion rate and hydrogen yield, the larger the SCR is the better, but in terms of system efficiency, it is most advantageous to operate at SCR 0.70 in Pressure Swing Adsorption (PSA) mode and SCR 0.80 in Pd membrane mode. Through this study, it was found that the optimal SCR in the reformer and the entire system including the reformer may be different, which indicates that the optimum operating point may be different depending on the change of the system configuration.

Large scale splitter-less FFD-SPLITT fractionation: effect of flow rate and channel thickness on fractionation efficiency (대용량 중력장 SPLITT Fractionation: 분획효율에 미치는 채널 두께와 유속의 영향)

  • Yoo, Yeongsuk;Choi, Jaeyeong;Kim, Woon Jung;Eum, Chul Hun;Jung, Euo Chang;Lee, Seungho
    • Analytical Science and Technology
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    • v.27 no.1
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    • pp.34-40
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    • 2014
  • SPLITT fractionation (SF) allows continuous (and thus a preparative scale) separation of micronsized particles into two size fractions ('fraction-a' and 'fraction-b'). SF is usually carried out in a thin rectangular channel with two inlets and two outlets, which is equipped with flow stream splitters at the inlet and the outlet of the channel, respectively. A new large scale splitter-less gravitational SF (GSF) system had been assembled, which was designed to eliminate the flow stream splitters and thus is operated by the full feed depletion (FFD) mode (FFD-GSF). In the FFD mode, there is only one inlet through which the sample is fed. There is no carrier liquid fed into the channel, and thus prevents the sample dilution. The effects of the sample-feeding flow rate, the channel thickness on the fractionation efficiency (FE, number % of particles that have the size predicted by theory) of FFD-GSF was investigated using industrial polyurethane (PU) latex beads. The carrier liquid was water containing 0.1% FL-70 (particle dispersing agent) and 0.02% sodium azide (used as bactericide). The sample loading rate was varied from about 4 to 7 L/hr with the sample concentration fixed at 0.01%. The GSF channel thickness was varied from 900 to $1300{\mu}m$. Particles exiting the GSF channel were collected and monitored by optical microscopy (OM). Sample recovery was monitored by collecting the fractionated particles on a $0.45{\mu}m$ membrane filter. It was found that FE of fraction-a was increased as the channel thickness increases, and FE of fraction-b was increased as the flow rate was increased. In all cases, the sample recovery has higher than 95%. It seems the new splitter-less FFD GSF system could become a useful tool for large scale separations of various types of micron-sized particles.

Effect of Trehalose on the Viability of Fluorescent Pseudomonas, Strain SSL3 (형광성 Pseudomonas, SSL3 균주의 생존율에 미치는 Trehalose의 효과)

  • Seong, Ki-Young;Ryu, Ok-Ran;Choi, Won-Yeol
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.4
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    • pp.292-301
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    • 2000
  • To make a better use of the beneficial bacterial inoculants in the agricultural practice, dry forms of bacterial fertilizer or pesticides are prepared with carrier materials. During the drying process of bacterial inoculant, most of the cells face a severe osmotic pressure and dehydration, and die off. Our study describes the effect of osmoprotectants such as trigonelline and trehalose on the survival of bacterial cells in high salt concentration and drying conditions. A fluorescent Pseudomonas, strain SSL3, used in this study, could grow in high salt concentration of upto 5% but the cells could not overcome the growth retardation at over 7% of salt concentration. The addition of trigonelline, even on small amount, in liquid medium containing 4% NaCl was detrimental to the cell. However, the addition of trehalose of upto 10 mM to the liquid medium containing 4% NaCl, enhanced cell growth. The cell growth was retarded when 150mM trehalose was added to the medium. Upon dry formulation of cells, trehalose was added. And the dry cells were inoculated into the soil to determine the effect of osmoprotectants on the survival of the cells. The survival of the cells, both in wet or dry soil, was improved by the addition of trehalose during the dry cell formulation. The positive effect of trehalose on the cell survival at $-20^{\circ}C$ and $-70^{\circ}C$ was oven more pronounced. The FTIR (Fourier transformation infra-red) spectroscopic analysis showed that the change of the 2nd amide group was reduced by adding trehalose to the medium containing 4% NaCl. These results suggest that trehalose can protect the cell membrane from dryness or high concentration of salt, thereby diminishing the sudden change of the protein structure of the cell membrane and, as a consequence, improving the cell survival.

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